| Background: Skin serves a variety of functions, the most important being as a barrier to the numerous hazards present in the environment. Other major functions include thermoregulation, fluid homeostasis, sensory detection and immune surveillance. The integration of the skin allows us to keep homeostasis and interact with our environment freely. Extensive skin loss from a variety of conditions such as burns is associated with significant functional morbidity and even loss of life. Full-thickness skin loss resulting from thermal injury presents several challenges including reconstruction of skin barrier function and prevention of sepsis and multisystem organ failure. Early excision of eschar with skin grafting minimizes these problem and reduces mortality rates. Autologous skin grafts provide ideal coverage but are in short supply on patients with large bums. CEA offer the potential to cover large surface defects from small amounts of skin. The production of CEA requires only small amounts (typically about 2x2cm2) of donor skin. The rapid in vitro growth of the keratinocytes isolated from the donor epidermis produces enough epithelium to cover the entire body surface within 3 weeks, corresponding to a 10,000-fold expansion of the original epithelial surface area. Several problems, however, are associated with clinical use of these grafts. The cultured cell sheets are initially fragile, susceptible to infection, and require special handling techniques. Especially, Graft production time, being both variable and somewhat lengthy, may be associated with difficulties in coordination of treatment schedules. In the contrast, epidermal cell suspension graft provide several advantages, especially immediate availability. Cells which uncultured or cultured to sub-confluence or to confluence all can be used in suspensions to close wound. The independence of time achieved in this way would allow for epithelial transplantation schedule in which both patient and cells are in the best possible condition. In this study, we added proper density sodium hyaluronate to the epidermal cell suspension in order to reduce the mobility of culture medium. Unfortunately, epithelial grafts alone do not provide an ideal permanent wound coverage, they provide only an epidermal component to full-thickness skin defects and often lack long-term durability. They have also been shown to blister or ulcerate withminimal traUma and highly susceptible to infection. In this stUdy, we use a collagenmembrane to act as a dermal scaffold to improve the quality of wound.Objective: l. St'Udy the proper density of SH matrix that not only can reduce themobility of cultUre medium, but also not influence the proliferation of keratinocy'tes;.2. Study the methods of preparing a collagen membrane stenuned from theskin of pig and investigate its effectiveness as a dermal scaffold.;3. StUdy the feasibility of the transplantaion of epidermal cell suspensionswith collagen membrane.Method8: l. Preparing the SH matrix: Keratinocyte cultUred in cultUre mediumcontalned SH with varian density for 48 or 72 hours, then with ELASA we determined theOD of the samples. By comparing the OD which can reflected the proliferation of cells, wefound out Whether the SH matrix influence the proliferation of keratinocyte. Thenintegrated considering the adhesion of the SH solution, we selected the proper density ofSH matrix.2. Preparing the collagen membrane: Pig hide collagen was dispersed in0.5M acetic acid, co-precipitated with chordroitin-6-sulfate and lyophilized for 48 hours toproduce a highly porous membrane. Cross-linking was occurred in 0.25% glutaraldehydefor 24 hours. The membrane was implanted beneath the flaPs on the dorsa... |
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